FR3090003A1 - New polymers and their use for the detection of ion fluxes - Google Patents

Abstract

The present invention relates to ion-sensitive polymers and their use for monitoring biological phenomena associated with ion fluxes, as well as organo-electrochemical transistors comprising such polymers. Figure for abstract: No figure.

Description

Description

Title of the invention: New polymers and their use for the detection of ion fluxes

The present invention relates to ion-sensitive polymers and their use for monitoring biological phenomena associated with ion fluxes, as well as organo-electrochemical transistors comprising such polymers.

Electrical activity is the basis of many primordial events in living systems, such as brain activity, heartbeat or hormonal secretion. This electrical activity is carried in particular by the transmembrane flow of ions of different types. These cellular signals are often recorded using probes that require genetic or chemical changes. In addition, the techniques used today to measure ion currents at the cellular level, in particular the “patch-clamp” technique, require a high level of expertise to be used and do not allow an ion current measurement to be carried out during more than a few minutes. It would be advantageous, in particular in order to be able to apply the technique to humans, to obtain unbiased signals, specific to a given type of ion, and to facilitate the use of the technique by recording intrinsic signals without using techniques as sophisticated as the "patch clamp". Extracellular microelectrode arrays (MEA) have been developed, allowing a non-invasive analysis of electrical phenomena at the cellular level for a long time, but on the one hand these systems are not specific for a type of ion, and on the other hand their signal to noise ratio remains low.

The detection of specific ion fluxes, that is to say fluxes of a given type of ion, is important for understanding the signaling of cells as well as their changes in the context of certain pathologies, genetic manipulations. , pharmacological or toxicological or during the maturation and / or differentiation of stem cells.

[0004] The detection of specific ion fluxes can also increase the specificity of biosensors using cells or micro-organs as sensors.

[0005] In addition, the development of organs on a chip, miniaturized devices using the combination of microfluidics and living cells to reproduce the complexity of human organs and ultimately of the whole organism, is today a major challenge for understanding of physiology and pathophysiology, development of new drugs or toxicity studies. In this context, it is important to be able to follow rigorously and non-invasively the specific ionic phenomena.

Detection of specific ions also allows rapid analysis of liquid samples from biological organisms, environmental liquid samples, and / or liquid samples taken during the monitoring of chemical processes.

Organo-electrochemical transistors (OECT), which are based on semiconductor polymers, base their operation on the movement of ions via non-invasive methods, allow amplification of signals without increasing noise and can give cell activity information. However, they cannot intrinsically discriminate between the different types of ions that participate in membrane ion fluxes, which prevents obtaining a more precise picture of the activity of the cells or micro-organs of interest.

To overcome this lack, selective electrodes vis-à-vis the ions have been designed. These electrodes are in the form of membranes selective for a given type of ion associated with conductive polymers. However, there is no material that is both electronic transducer and sensitive to ions, which would allow in particular to improve the specificity for a given type of ion, and to reach large areas by repetitive printing.

The inventors have developed in this context new polymers and polymer complexes which are both electronic conductors and specific for target ions, as well as conductive inks comprising said polymers or complexes. The polymers are either electronic conductors in themselves, or they form a conductive complex in admixture with an electronic conductive polymer. The inks obtained are particularly suitable for use in the manufacture of electrodes and transistors for recording signals at the cellular level. The use of these polymers and inks in transistors makes it possible to combine:

- the advantages of polymers and inks, in particular the improvement of the specificity for a type of ions, and the repetitive printing of micrometric units on large surfaces and

- the advantages of transistors, in particular the reduction of detection thresholds via amplification and noise reduction.

The present invention relates firstly to the use of at least one polymer comprising at least one unit of formula (I)

[Chem.l] —A— ^: LIB

In which

A is a polymerizable monomer,

L is a spacer arm, and

B is a chemical group capable of complexing or chelating at least one ion chosen from the group consisting of K ⁺ , Na ⁺ , Ca ²⁺ and Zn ²⁺ , in a process for coating an electrode or manufacturing a transistor organo-electrochemical.

It also relates to a conductive ink comprising at least one polymer comprising at least one unit of formula (I)

[Chem.2] • L i B

In which A is a polymerizable monomer, L is a spacer arm, and B is a chemical group capable of complexing or chelating at least one ion chosen from the group consisting of K ⁺ , Na ⁺ , Ca ²⁺ and Zn ^{2 +} , in which either the polymer comprising at least one unit of formula (I) is an electronic conductive polymer, or the polymer comprising at least one unit of formula (I) is non-electronic conductor and is in the form of a mixture with another electronic conductive polymer.

It also relates to an organo-electrochemical transistor comprising as a semiconductor film a film comprising at least one polymer comprising at least one unit of formula (I)

[Chem.3] —j -— A- ^ | -

L I B

In which A is a polymerizable monomer, L is a spacer arm, and B is a chemical group capable of complexing or chelating at least one ion chosen from the group consisting of K ⁺ , Na ⁺ , Ca ²⁺ and Zn ^{2 +} .

It also relates to the use of such a transistor for the detection of at least one flux of an ion chosen from the group consisting of K ⁺ , Na ⁺ , Ca ²⁺ and Zn ²⁺ at the level of a cell or set of cells.

It also relates to the use of such a transistor for the detection of the presence of at least one ion chosen from the group consisting of K ⁺ , Na ⁺ , Ca ²⁺ and Zn ²⁺ in a liquid sample from of a biological organism, in an environmental liquid sample, or in a liquid sample taken during the monitoring of a chemical process.

It also relates to a polymer comprising at least one unit of formula (I) [Chem.4] —A— ' ^: L

I

B

In which A is a polymerizable monomer, L is a spacer arm, and B is a chemical group capable of complexing or chelating at least one ion chosen from the group consisting of K ⁺ , Na ⁺ , Ca ²⁺ and Zn ^{2 +}

Finally, it relates to a monomer of formula (II)

[Chem.5]

A — L — 8

In which A is a polymerizable monomer, L is a spacer arm, and B is a chemical group capable of complexing or chelating at least one ion chosen from the group consisting of K ⁺ , Na ⁺ , Ca ²⁺ and Zn ^{2 +} , in which the monomer of formula (II) is chosen from the group consisting of

- 4-vinylbenzyl (sulfonyl) -4 '- (benzo-15-crown-5) -ylamine 1,

- 4-vinylbenzyl (sulfonyl) -4 '- (benzo-18-crown-8) -ylamine 2,

- 4-vinylbenzyl-4 '- (methyl-15-crown-5) -methylether 4,

- 4-vinylbenzyl-4 '- (methyl-18-crown-8) -methylether 5,

-la (2,3-dihydrothieno [3,4-b] [1,4] dioxin-2-yl) -4 '- (benzo-15-crown-5) -methylamine 6, -la (2,3- dihydrothieno [3,4-b] [1,4] dioxin-2-yl) -4 '- (benzo-18-crown-6) -methylamine 7, -la (2,3-dihydrothieno [3,4-b ] [l, 4] dioxin-2-yl) -N, N-bis ((pyridin-2-yl) methyl) methanamine 8,

- (2,3-dihydrothieno [3,4-b] [1,4] dioxin-2-yl) -2- (methyl-15-crown-5) -methylether 9, and - le (2,3- dihydrothieno [3,4-b] [1,4] dioxin-2-yl) -2- (methyl-18-crown-6) -methyl ether 10.

Other features and advantages will appear on reading the description below. The figures are presented for information only and are in no way limitative of the invention.

[Fig-1] presents the synthesized inks: from left to right, PEDOT ink:

PSTESI_80 -co- PS15-crown-5 (l) _20, PEDOT ink: PSTESI_85 -coPS15-crown-5 (1) _15, PEDOT ink: PSTESI_85 -co- PS18-crown-6 (2) _15 and PEDOT ink: PSTESI_85 -co- PS18-crown-6 (2) (250 KDa) _15 (a); two PEDOT-based inks in their doped and dedoped states (electrolyte O, 1M NaCl, V = -0.75mV) (b and c): PEDOT: Commercial PSS, transparent in its oxidized state (b left) and blue in its reduced state (b right); PEDOT: PSTFSI-85: PSP15Cr5SI-15, transparent in the oxidized state (c left) and blue in the reduced state (c right).

[Fig.2] presents the electrochromic characterization of the ink dedoping time

PEDOT: PSTFSI-85: PSP15Cr5SI-15 (a) and the characterization in electro-impedance spectroscopy (EIS) of PEDOT ink: PSTFSI-85: PSP15Cr5SI-15 (b).

[Fig.3] shows the morphology of β-clonal cells (line INS-832/13) cultivated on coverslips with / without the thin layer of the film of PEDOT-based inks stabilized by the electrolyte polymers containing the fractions F6, F7 and F8 sensitive ions described in Table 2.

[Fig.4] presents the effect of thapsigargine on β-clonal cells (line INS832 / 13) cultured on coverslips with / without a thin layer of film of an ink of the PEDOT complex: polymer according to the invention, the polymer according to the invention being a copolymer between STFSI and monomer 2, in mass proportions 85/15 (PEDOT: PSTFSI 85-co-PS18-crown-6). (a) and (b) are images on lamellae without a thin layer of ink according to the invention, (c) and (d) are images with a thin layer of ink according to the invention, (a) and (c) are images in the absence of thapsigargine, (b) and (d) are images in the presence of thapsigargine, (e) represents the quantification of the percentage of dead cells for each condition.

[Fig.5] presents (a) the electrochemical characterization of the monomer 6 compared to that of the EDOT monomer, (b) the electrochemical characterization of the various copolymers of the monomer 6 with the EDOT, in the order of the curves ( starting from the top at the level of 0.0 on the abscissa) with EDOT / monomer 6 ratios of 2/1, 3/1, 2/3 and 1/1.

[Fig.6] presents (a) a scanning electron microscopy image of the copolymer

EDOT / monomer 6 with a 3/1 ratio, (b) a scanning electron microscopy image of a 20 micron film formed by electropolymerization of monomer 6.

[Fig.7] shows a diagram of the manufacture and characterization of an OECT according to the invention.

[Fig.8] represents the physical characterization of an OECT according to the invention. The curve with dots (left axis) is the transfer curve showing the “on” state (at a grid voltage of -0.2V) and the “off” state (at a grid voltage of +0 , 75 V) of the electrochemical transistor (VD = -0.4V). The curve with squares (axis on the right) is the transconductance of the electrochemical transistor showing a maximum of amplification at low voltage (Vg = + 0.2 V) and with a magnitude similar to those of the state of the art ( VD = -0.4V).

The present invention relates first of all to the use of at least one polymer comprising at least one unit of formula (I)

[Chem.6] + * +; L

I

In which

A is a polymerizable monomer,

L is a spacer arm, and

B is a chemical group capable of complexing or chelating at least one ion chosen from the group consisting of K ⁺ , Na ⁺ , Ca ²⁺ and Zn ²⁺ , in a process for coating an electrode or manufacturing a transistor organo-electrochemical.

Thus, the invention covers a method of coating an electrode or manufacturing an organo-electrochemical transistor comprising at least one step of using at least one polymer comprising at least one unit of formula (I )

[Chem.7] • L I B

In which

A is a polymerizable monomer,

L is a spacer arm, and

B is a chemical group capable of complexing or chelating at least one ion chosen from the group consisting of K ⁺ , Na ⁺ , Ca ²⁺ and Zn ²⁺ .

These ions are important ions in electrophysiology. Indeed, the cations K ⁺ , Na ⁺ and Ca ²⁺ are notably known to carry electric currents which cross the cell membrane. The Zn ²⁺ cation is particularly important in the context of diabetes, since the pancreatic beta cells release molar quantities of Zn ²⁺ and insulin in a determined ratio (2 Zn ²⁺ for 6 molecules of insulin) . Monitoring the released Zn ²⁺ therefore allows monitoring of the amount of released insulin, which is an important parameter in the monitoring of diabetes, in particular to adapt the treatment administered.

By "polymerizable monomer" means a monomer comprising at least one reactive function capable of forming a covalent bond with another monomer. The polymerizable monomers according to the invention can form electronic conductive polymers by homopolymerization or by copolymerization with at least one other monomer, preferably another monomer according to the invention. For example, mention may be made of the 3,4-ethylenedioxythiophene monomer (EDOT). Alternatively, the polymerizable monomer according to the invention can form a non-conductive polymer in itself by homopolymerization or by copolymerization with at least one other monomer, but form a conductive complex in the presence of another electronic conductive polymer. For example, mention may be made of the styrene and styrene sulfonate monomers, in particular sodium styrene sulfonate, which can be used as stabilizers during the oxidative polymerization of EDOT in water to form a conductive complex such as PEDOT: PSS which is a mixture of poly (3,4-ethylenedioxythiophene) and sodium poly (styrene sulfonate).

In one embodiment, the polymerizable monomer A is chosen from the group consisting of styrene, styrene sulfonate, in particular sodium styrene sulfonate, and 3,4-ethylenedioxythiophene.

By "spacer arm" means a succession of atoms covalently linked. Typically, the spacer arm is a group comprising at least one atom of carbon, hydrogen, nitrogen, and / or oxygen. For example, the chain of atoms of the spacer arm connecting the polymerizable monomer A and the chemical group capable of complexing or chelating a B ion comprises from 1 to 10 atoms, preferably from 1 to 5 atoms, in particular from 1 to 3 atoms. In particular, the spacer arm L can be an oxygen atom, an alkyl group, an ether group, an amine group such as a secondary or tertiary amine, an alkylamine group, an amide group, an ester group, a cycloalkyl group, a heterocyclic group or a ketone group. Preferably, the spacer arm is chosen from the group consisting of an amine group, in particular NH, an alkyl group, in particular CH ₂ , an ether group, in particular CH ₂ -O-CH ₂ , and an alkylamine group, in particular CH ₂ -NH. In the case where the spacer arm is not symmetrical, it can be placed both in one direction and in the other between the polymerizable monomer A and the chemical group capable of complexing or chelating a B ion. The spacer arm can also include several groups as listed above covalently linked together. For example, the spacer arm can be an alkylamine group, an alkylester group, an alkylamide group, or an alkylketone group.

By "alkyl group" means a saturated hydrocarbon group, linear or branched, comprising from 1 to 4 carbon atoms. Among the examples of alkyl groups, mention may be made of methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl and iso-butyl groups, preferably methyl and ethyl groups.

By “ether group” is meant two alkyl groups covalently linked to each other by an oxygen atom. For example, mention may be made of the group ch ₂ -o-ch ₂ .

By "amine group" means an NH group or a Nalkyl group.

By "amide group" means a group C (= O) -NH.

By "ester group" means a group C (= O) -O.

By "cycloalkyl group" means a hydrocarbon ring, saturated, unsaturated or aromatic, preferably saturated, and comprising from 1 to 10 carbon atoms. The cycloalkyl group may in particular be a cyclopropane, a cyclobutane, a cyclopentane, a cyclohexane, a cycloheptane, a cyclooctane, a cyclobutene, a cyclopentene, a cyclohexene, or a phenyl.

By "heterocyclic group" is meant a hydrocarbon, saturated, unsaturated or aromatic ring, comprising from 1 to 10 members, and interrupted by at least one heteroatom chosen from the group consisting of oxygen, nitrogen and sulfur. Preferably, the heterocyclic group comprises 5 or 6 links. The heterocyclic group can in particular be a furan, a tetrahydrofuran, a thiophene, a pyrrole, a pyridine, a pyrane, an oxazine, a thiazine, a pyrimidine or a piperazine. The heterocyclic group may possibly comprise several cycles, for example it may be a bicyclic group.

By "ketone group" means a group C (= O).

By "chemical group capable of complexing or chelating at least one ion" means a chemical group whose structure allows a non-covalent interaction with at least one ion. Preferably, the non-covalent interaction between the chemical group and the at least one ion is specific, that is to say that the affinity of the chemical group for the target ion is at least 2 times, preferably at least 10 times, greater than that for another ion in the list, preferably that for any other ion in the list.

For example, B can be chosen from crown ethers, cyclic ionophores such as valinomycin and nonactin, and the di (2-picolyl) amine (or Bis (2-pyridylmethyl) amine) group. Preferably, B is chosen from crown ethers and the di (2-picolyl) amine group.

By "crown ethers" is meant cyclic oligomers of ethylene oxide, optionally substituted, known for their ability to interact with cations. Among the examples of crown ethers, mention may be made of ether 12-crown-4, ether 15-crown-5, ether 18-crown-6 and ether 21-crown-7, optionally substituted, in particular substituted with at least one benzyl. Preferably, the crown ether is 15-crown-5 ether or 18-crown-6 ether, optionally substituted, in particular by a benzyl. The sulfur analogs of oxygenated crown ethers (crown thioethers) are also included in the scope of the term "crown ethers" according to the invention. As such, mention may be made in particular of 1,5,9,13-tetrathiacyclohexadecane. The nitrogen analogs of oxygenated crown ethers are also included in the scope of the term "crown ethers" according to the invention, as well as the mixed analogs of aza crown ether. As such, there may be mentioned respectively hexaazacyclooctadecane, 1,4,7,10-tetraazacyclododecane (nitrogen analogues) and diaza-18-crown-6 (mixed analog).

In one embodiment, the ion is the potassium ion K ⁺ and B is chosen from the group consisting of 18-crown-6 ether (1,4,7,10,13,16-hexaoxacyclooctadecane ), optionally substituted, valinomycin and nonactin.

In one embodiment, the ion is the sodium ion Na ⁺ and B is the ether 15-crown-5 (1,4,7,10,13-pentaoxacyclopentadecane), optionally substituted.

In one embodiment, the ion is the zinc ion Zn ²⁺ and B is the di (2-picolyl) amine group, optionally substituted.

The polymer comprising at least one unit of formula (I) is either an electronic conductive polymer, or it is non-electronic conductor and is used in the form of a mixture with another electronic conductive polymer. In this second case, the polymer according to the invention is advantageously a stabilizer during the synthesis of the electronic conductive polymer with which it is mixed. The mixture with an electronic conductive polymer is also designated “conductive complex” in the present invention.

By "electronic conductive polymer" means a polymer which alternately has single and multiple bonds, capable of conducting the electrons. It can be a conductive or semi-conductive polymer, possibly doped to increase its conduction properties.

The method of coating an electrode or manufacturing an organoelectrochemical transistor can be any method conventionally implemented in this field. The electrode may be an electrode for an organo-electrochemical transistor.

In one embodiment, the polymer comprising at least one unit of formula (I) is used in the form of an ink, that is to say it is included in a conductive ink.

The conductive ink, the conductive complex or the conductive polymer comprising at least one motif of formula (I) can be deposited on the electrode or on the substrate of the transistor by any technique known in the art, in particular by centrifugation (spin coating) or by inkjet printing.

The invention also relates to a conductive ink comprising at least one polymer comprising at least one unit of formula (I)

[0066]

[Chem. 8] + H; L

I

B

In which A, L and B are as defined above, in which either the polymer comprising at least one unit of formula (I) is an electronic conductive polymer, or the polymer comprising at least one unit of formula ( I) is non-electronic conductor and is in the form of a mixture with another electronic conductive polymer.

A conductive ink according to the invention is a complex mixture of compounds comprising at least one polymer comprising at least one unit of formula (I) and at least one additive, for example a solvent or a formulating agent. The solvent can in particular be water. A conductive ink according to the invention has physicochemical properties such as its wettability and / or its viscosity in particular which allow it to be deposited or printed. The printing of the conductive ink according to the invention produces an electronic conductive printed object. The conductive ink according to the invention may include additives conventionally used in bioelectronics for obtaining films with the best properties. For example, the ink may contain at least one compound to improve its conductivity, for example a co-solvent such as dimethyl sulfoxide (DMSO) or ethylene glycol (EG), at least one compound to increase the adhesion to the surface , for example the 3-glycidoxypropyltrimethoxysilane crosslinker or the divinylsulfone crosslinker (DVS), at least one compound for increasing the stability of the conductive ink film in an aqueous medium, for example the 3-glycidoxypropyltrimethoxysilane crosslinker or the divinylsulfone crosslinker ( DVS), and / or at least one compound for improving wetting on the substrate, for example a surfactant from the class of Zonyl® (polytetrafluoroethylene resins) or 4-dodecylbenzene sulfonic acid (DBSA).

This ink can for example be an ion-sensitive ink based on PEDOT obtained by stabilizing an aqueous dispersion of EDOT monomers with at least one polymer comprising at least one unit of formula (I) as defined above, in particular at least one copolymer of at least one of the 4-vinylbenzyl (sulfonyl) -4 '- (benzo-15-crown-5) -ylamine 1, 4-vinylbenzyl (sulfonyl) -4' - (benzo-18-crown) monomers -8) -ylamine 2 and N (4-vinylbenzyl) (pyridin-2-yl) -N - ((pyridin-2-yl) methyl) methylamine 3, with styrene (trifluoromethanesulfonyl) imide (STFSI), then by performing the oxidative polymerization of said dispersion. This provides ion sensitivity to a conductive PEDOT-based ink.

During their use, the inks according to the invention can be formulated, deposited in the form of films using various known techniques. The technique used can be adapted according to the viscosity of the ink and can be 'spin coating', 'doctor blade', 'slot die' or 'spray coating'. In addition, the inks can be characterized by different electrochemical methods, in particular for the determination of their ionic sensitivity.

The invention also relates to an organo-electrochemical transistor comprising as semiconductor film a film comprising at least one polymer comprising at least one unit of formula (I)

[Chem.9]

L i

B

In which A, L and B are as defined above. This transistor has the advantage of being directly ion-specific without requiring the combination of an ion-specific membrane and a conductive polymer. The manufacturing process of the transistor is therefore simplified since the conductive polymer or the conductive complex fulfills the double role of ion selectivity and conduction. The transistors according to the invention can advantageously be used as ion-specific sensors for recordings in electrophysiology in particular, for research applications and in the biomedical field.

In one embodiment, the semiconductor film of the transistor according to the invention is obtained by printing or depositing a conductive ink according to the invention.

Either the polymer comprising at least one unit of formula (I) included in the film of the transistor according to the invention is an electronic conductive polymer, or it is non-electronic conductor and is in the form of a mixture with another electronic conductive polymer.

The use of a transistor according to the invention for the detection of at least one flux of an ion chosen from the group consisting of K ⁺ , Na ⁺ , Ca ²⁺ and Zn ²⁺ at the level of another cell or set of cells is another object of the invention. The cell or set of cells can either be in vivo or be in vitro, for example within a cell culture, preferably in vitro. The cell or the set of cells can in particular be on the transistor. The transistor can be used for the detection of specific ion fluxes during physiological, physiopathological (functional explorations and diagnostics), pharmacological, toxicological or organ-on-chip investigations. The transistor can also be used for the detection of specific ion fluxes during the maturation and / or differentiation of stem cells, for example to monitor their differentiation and then improve the differentiation protocols or when using differentiated stem cells as a model. genetic disease. The transistor can also be used according to the invention for the detection of specific ionic fluxes at the level of a bio-sensor using cells, for example clonal cells, primary cells or stem cells, or microorganisms as sensors. In a particular embodiment, and for legal or ethical reasons, stem cells are to the exclusion of human embryonic stem cells. Finally, the transistor can be used according to the invention for the detection of Zn ²⁺ co-secreted with insulin at the beta-pancreatic cells, in particular for monitoring and / or quantifying the secretion of insulin in a patient with diabetes.

The use of a transistor according to the invention for the detection of the presence and / or the quantification of at least one ion chosen from the group consisting of K ⁺ , Na ⁺ , Ca ²⁺ and Zn ²⁺ in a liquid sample from a biological organism, in an environmental liquid sample, or in a liquid sample taken during the monitoring of a chemical process is another object of the invention.

The liquid sample to be analyzed can for example be a liquid sample comprising cells, an organ, a biological organism, an environmental liquid sample or a liquid sample taken during the monitoring of a chemical process.

Insofar as the conductive polymers, conductive complexes and conductive inks according to the invention can be deposited on flexible supports, the transistors according to the invention make it possible to overcome the rigidity of the metal electrodes which is a major concern. in interfacing between electrodes and biological material, in particular in vivo.

In addition, they make it possible to detect ionic fluxes by overcoming the problems inherent in optical methods such as photobleaching, heating or bias by the use of organic probes or those resulting from genetic modifications.

The organoelectrochemical transistor according to the invention can be manufactured by any technique known in the art for manufacturing OECTs. The conductive ink according to the invention can for example be deposited on a glass or plastic substrate such as polyethylene terephthalate (PET).

The invention also relates to a polymer comprising at least one unit of formula (I)

[0083]

[Chem. 10] + H; L I B

In which A is a polymerizable monomer, L is a spacer arm, and B is a chemical group capable of complexing or chelating at least one ion chosen from the group consisting of K ⁺ , Na ⁺ , Ca ²⁺ and Zn ^{2 +} . The polymer according to the invention can only comprise units of formula (I) (homopolymer), or a combination of such units with at least one other unit (copolymer). These other units can advantageously be EDOT units, STESI units, styrene units, styrene sulfonate units, in particular sodium styrene sulfonate, or units of formula (I) with A, L and / or B different from those of first motif.

The polymer according to the invention may in particular be a copolymer between a 4-vinylbenzyl (sulfonyl) -4 '- (benzo-15-crown-5) -ylamine 1, 4-vinylbenzyl (sulfonyl) -4'- monomer (benzo-18-crown-8) -ylamine 2 or N (4-vinylbenzyl) (pyridin-2-yl) -N - ((pyridin-2-yl) methyl) methylamine 3 and the monomer STESI. In this case, it is a non-conductive polymer but forms a conductive complex when it is mixed with PEDOT or a derivative or analog thereof. The polymer according to the invention acts in this case as a stabilizer during the synthesis of PEDOT or of the derivative or analog thereof, in particular when the latter is polymerized oxidatively in an aqueous (dispersed) medium.

The polymer according to the invention may in particular be a copolymer between a 4-vinylbenzyl-4 '- (methyl-15-crown-5) -methylether 4 or 4-vinylbenzyl-4' - (methyl-18-crown) monomer -8) -methylether 5 and the monomer STESI. In this case, it is a non-conductive polymer but forms a conductive complex when it is mixed with PEDOT or a derivative or analog thereof.

The polymer according to the invention may alternatively be a homopolymer of a monomer chosen from the group consisting of monomers (2,3-dihydrothieno [3,4-b] [1,4] dioxin-2-yl) - 4 '- (benzo-15-crown-5) -methylamine 6, (2,3-dihydrothieno [3,4-b] [1,4] dioxin-2-yl) -4' - (benzo-18-crown -6) -methylamine 7, (2,3-dihydrothieno [3,4-b] [1,4] dioxin-2-yl) -N, N-bis ((pyridin-2-yl) methyl) methanamine 8, (2,3-dihydrothieno [3,4-b] [1,4] dioxin-2-yl) -2- (methyl-15-crown-5) -methyl ether 9, and (2,3-dihydrothieno [3, 4-b] [1,4] dioxin-2-yl) -2- (methyl-18-crown-6) -methylether 10, in particular a homopolymer of monomer 6. It is in this case a conductive polymer electronic.

The polymer according to the invention may alternatively be a copolymer between a monomer chosen from the group consisting of monomers 6, 7, 8, 9 and 10 and an EDOT monomer, in particular a copolymer between monomer 6 and an EDOT monomer. In this case, it is an electronic conductive polymer.

In the case of copolymers involving units which are not of formula (I), the ratios by mass of units of formula (I) / other units are advantageously included in the range 5/95 to 50/50.

The polymer according to the invention can have a variable chain length over a wide range, in particular in the range conventionally used for the use of PEDOT or PEDOT: PSS as a conductive polymer or conductive complex respectively. When, in formula (I), A is styrene or styrene sulfonate, in particular sodium styrene sulfonate, the polymer according to the invention typically has a molar mass of between 100 and 300 kg / mol.

The polymers according to the invention can be obtained by polymerization of monomers of formula (II)

[Chem. 11]

A-L-B

In which A, L and B are as defined above, or by copolymerization of such monomers with other monomers. The polymerization can be carried out by any technique known in the field for polymerizing the monomers of formula A. For example, the electronic conductive polymers according to the invention which are such that A is PEDOT and are derivatives or analogs of PEDOT can be obtained by radical oxidative polymerization, or by electropolymerization. The non-conductive polymers according to the invention which are such that A is styrene sulfonate and are derivatives or analogs of polystyrene sulfonate can be obtained by radical polymerization, advantageously by radical polymerization controlled via RAFT (radical polymerization controlled by reversible chain transfer by additionfragmentation) or NMP (radical polymerization in the presence of nitroxides).

In one embodiment, the unit of formula (I) included in the polymer according to the invention is chosen from the group consisting of:

- 4-vinylbenzyl (sulfonyl) -4 '- (benzo-15-crown-5) -ylamine 1, - 4-vinylbenzyl (sulfonyl) -4' - (benzo-18-crown-8) -ylamine 2, - N- (4-vinylbenzyl) (pyridin-2-yl) -N - ((pyridin-2-yl) methyl) methylamine 3, - 4-vinylbenzyl-4 '- (methyl-15-crown-5) -methylether 4, - 4-vinylbenzyl-4 '- (methyl-18-crown-8) -methylether 5, -la (2,3-dihydrothieno [3,4-b] [1,4] dioxin-2- yl) -4 '- (benzo-15-crown-5) -methylamine 6, -la (2,3-dihydrothieno [3,4-b] [1,4] dioxin-2-yl) -4' - ( benzo-18-crown-6) -methylamine 7, -la (2,3-dihydrothieno [3,4-b] [1,4] dioxin-2-yl) -N, N-bis ((pyridin-2- yl) methyl) methanamine 8,

- (2,3-dihydrothieno [3,4-b] [1,4] dioxin-2-yl) -2- (methyl-15-crown-5) -methyl ether 9, and

- (2,3-dihydrothieno [3,4-b] [1,4] dioxin-2-yl) -2- (methyl-18-crown-6) -methylether 10.

Preferably, the motif of formula (I) is chosen from the group consisting of: - 4-vinylbenzyl (sulfonyl) -4 '- (benzo-15-crown-5) -ylamine 1, - 4- vinylbenzyl (sulfonyl) -4 '- (benzo-18-crown-8) -ylamine 2, - 4-vinylbenzyl-4' - (methyl-15-crown-5) -methyl ether 4, - 4-vinylbenzyl-4 '- (methyl-18-crown-8) -methylether 5, -la (2,3-dihydrothieno [3,4-b] [1,4] dioxin-2-yl) -4' - (benzo-15- crown-5) -methylamine 6, -la (2,3-dihydrothieno [3,4-b] [1,4] dioxin-2-yl) -4 '- (benzo-18-crown-6) -methylamine 7 , -la (2,3-dihydrothieno [3,4-b] [1,4] dioxin-2-yl) -N, N-bis ((pyridin-2-yl) methyl) methanamine 8,

- (2,3-dihydrothieno [3,4-b] [1,4] dioxin-2-yl) -2- (methyl-15-crown-5) -methylether 9, and - le (2,3- dihydrothieno [3,4-b] [1,4] dioxin-2-yl) -2- (methyl-18-crown-6) -methyl ether 10. The invention also relates to an electronic conductive complex comprising a polymer according to the invention, preferably a non-conductive polymer according to the invention, and an electronic conductive polymer. The electronic conductive polymer in the electronic complex can be, for example, PEDOT or a derivative or analog thereof. A PEDOT derivative can in particular be obtained by replacing at least certain EDOT monomers with substituted EDOT monomers, in particular at the level of at least one of the carbon atoms of the ethylenedioxy group of the EDOT monomer, with at least one chemical group. A PEDOT analog can in particular be obtained by replacing at least certain EDOT monomers with monomers in which the ethylenedioxy group is replaced by a propylenedioxy group, optionally substituted, or by replacing one or more of the oxygen oxygen atoms. at least some EDOT monomers with nitrogen, selenium or sulfur atoms.

The polymers and / or the electronic conductive complexes according to the invention combine the properties of conduction and ionic sensitivity. Due to this combination, their use can advantageously be envisaged in conductive inks, in organo-electrochemical transistors, or in any other system for detecting and / or quantifying said ions, in order for example to determine their concentration in analytes.

The invention finally relates to a monomer of formula (II),

[Chem. 12]

A-L-B

In which A is a polymerizable monomer, L is a spacer arm, and B is a chemical group capable of complexing or chelating at least one ion chosen from the group consisting of K ⁺ , Na ⁺ , Ca ²⁺ and Zn ^{2 +} .

Preferably, the monomer according to the invention is chosen from the group consisting of:

- 4-vinylbenzyl (sulfonyl) -4 '- (benzo-15-crown-5) -ylamine 1,

- 4-vinylbenzyl (sulfonyl) -4 '- (benzo-18-crown-8) -ylamine 2,

- 4-vinylbenzyl-4 '- (methyl-15-crown-5) -methylether 4,

- 4-vinylbenzyl-4 '- (methyl-18-crown-8) -methylether 5,

- (2,3-dihydrothieno [3,4-b] [1,4] dioxin-2-yl) -4 '- (benzo-15-crown-5) -methylamine 6, -la (2,3- dihydrothieno [3,4-b] [1,4] dioxin-2-yl) -4 '- (benzo-18-crown-6) -methylamine 7, -la (2,3-dihydrothieno [3,4-b ] [l, 4] dioxin-2-yl) -N, N-bis ((pyridin-2-yl) methyl) methanamine 8,

- (2,3-dihydrothieno [3,4-b] [1,4] dioxin-2-yl) -2- (methyl-15-crown-5) -methyl ether 9, and

- (2,3-dihydrothieno [3,4-b] [1,4] dioxin-2-yl) -2- (methyl-18-crown-6) -methylether 10.

[0102]

The examples below are intended to illustrate the invention without limiting its scope.

Examples

Example 1: Synthesis of monomers according to the invention

Synthesis of monomers 1 and 2

Step 1- Formation of 4-styrenesulfonyl chloride:

In a three-necked flask, flamed and dried beforehand, 60 ml of dry and degassed acetonitrile were introduced. Then, oxalyl chloride (5.91 g; 46.6 mmol; 1.2 eq.) And dimethylformamide (DMF) were added. This solution was stirred intensely at room temperature to dissolve the reagents and promote the formation of the Vilsmaier-Haack complex for 4-5 hours. Once the characteristic yellow color was obtained stably, sodium 4-styrenesulfonate (8.0 g; 38.8 mmol; 1.0 eq.) Was added to the reaction medium under an inert atmosphere and at room temperature. After 24 hours of reaction, the precipitated salt was separated by filtration and the reaction mixture was used in the second step.

Step 2- Synthesis of 4-vinylbenzyl (sulfonyl) -4 '- (benzo-15-crown-5) -ylamine (Sphl5cr5SI):

Anhydrous acetonitrile (V = 50 mL), triethylamine (4.29 g; 30.2 mmol; 3 eq.) And 4'- were added to a dry, flamed three-necked flask beforehand. aminobenzo-15-crown-5 (3.00 g, 10.1 mmol, 1 eq.). This mixture was stirred for one hour. Meanwhile, the 4-styrenesulfonyl chloride solution (from step 1) was cooled to 0 ° C. Then the 4’-aminobenzo-15-crown-5 solution was added to the latter in vacuo. After the addition, the reaction mixture was stirred at room temperature for 16 hours. The solvent of the reaction medium was evaporated under reduced pressure and the residue obtained was dissolved in dichloromethane, then successive extractions with potassium carbonate were carried out, followed by washing with an IM solution of hydrochloric acid. Finally, the organic solvent was evaporated and the residue obtained dried under vacuum for 16 h at 45 ° C. The product was obtained in the form of a viscous solid of brown color (yield (Yield) = 54%).

0-NMR (400 MHz; CDC1 ₃ ; 298K): 7.73 ppm (d, 2H); 7.57 (d, 2H); 6.79 (q, 1H); 6.71 (d, 1H); 6.29 (s, 1H); 6.21 (d, 1H); 5.95 (d, 1H); 5.38 (d, 1H); 3.85 (m, 4H); 3.74 (dt, 4H); 3.67-3.63 (m, 8H); ¹³ C-NMR (75 MHz; DMSO-d6; 298K): ppm 144.21; 139.55; 135.57; 126.43; 125.87; 126.36-122.06-117.77-113.47 (qCF ₃ ); 116.55; FTIR (ATRcm-1): = 1400, 1329, 1281, 1195, 1172, 1159, 1140, 1090, 1058, 844, 776, 735, 655; m / z (ESI-HRMS) 313.9779 ([M] C9H7F3NO4S2 corresponds to 313.9769).

The monomer 2 was synthesized in the same way by replacing the 4’-aminobenzo-15-crown-5 with the 4’-aminobenzo-18-crown-6.

Synthesis of monomers 4 and 5

The monomers 4 and 5 were synthesized in a single step consisting of etherification at low temperature in the presence of sodium hydride.

A solution of 2-hydroxymethyl-15-crown-5 (1.55 g; 5.9 mmol) in 15 ml of DMF was added dropwise to a solution of sodium hydride (0.307 g; 7 , 67 mmol) in DMF under an inert atmosphere at 0 ° C. After thirty minutes at this temperature, 4-vinylbenzyl chloride (1.109 mL; 7.08 mmol) was added. The reaction mixture was allowed to stir at 0 ° C for 2 hours before allowing it to return to room temperature. The reaction was stopped by adding water (70 mL). The organic phase was extracted several times with ethyl acetate, and the various combined organic phases were dried with sodium sulfate and concentrated in vacuo. The final product 4 (a colorless oil, 2.4 g; Yield = 93%) was purified by liquid chromatography with silica gel as stationary phase and a mixture of dichloromethane / diethyl ether (10: 1) as mobile phase.

Ή-NMR (CDC1 ₃ , 400 MHz): ô (ppm): 7.30 (dd, 2H); 7.21 (dd, 2H); 6.63 (q, 1H); 5.68 & 5.54 (d, 2H); 4.46 (s, 2H); 3.59 (m, 4H); 3.59 (m, 19 H). ¹³ C-NMR (CDC1 ₃ , 150 MHz): ô (ppm): 136.9; 135.8; 126.7; 125.1; 112.7; 77.7; 69.5; 69.4.

The monomer 5 was synthesized in the same way by replacing 2-hydroxymethyl-15-crown-5 with 2-hydroxymethyl-18-crown-6.

Synthesis of Monomers 6 and 7

The synthesis of monomers 6 and 7 was carried out in a single nucleophilic substitution step between chloromethyl-EDOT and 4'-aminobenzo-15-crown-5 or 4'-aminobenzo-18-crown-6 in presence of potassium carbonate.

In a 100 ml three-necked flask, under an inert atmosphere, were successively added chloro-methyl-EDOT (0.3436 g; 1.71 mmol; 1 eq.), 4'-aminobenzo-15-crown -5 (0.5 g; 1.71 mmol, leq.) And sodium carbonate (0.561 g; 5.29 mmol; 3eq.). The whole was dissolved in dry acetonitrile (40 mL) using a magnetic stirrer and heated to reflux (70 ° C) for 48 hours under an inert atmosphere. After returning to room temperature, the solvent was evaporated in vacuo and the crude product obtained was dissolved in 30 ml of dichloromethane. Its purification was carried out by liquid-liquid extraction with water (3x10 mL). The organic phase was then dried with magnesium sulfate. After filtration and evaporation of the solvent and drying under vacuum at 45 ° C for 24 hours, the pure monomer 6 was obtained in the form of a brown viscous oil (0.703 g, yield = 90%).

Ή-NMR (400 MHz; CDC1 ₃ ; 298K): ppm 6.71 (d, 1H); 6.35 (m, 2H); 6.27 (d, 1H); 6.20 (dd, 1H); 4.48 (s, 16H); 4.35 (m, 1H); 4.28-4.12 (m, 2H); 4.05 (m, 4H); 3.87 (m, 4H); 3.74 (s, 8H); 3.71-3.63 (m, 2H). ¹³ C-NMR (75 MHz; CDC13; 298K): / / ppm: 150.7; 142.10; 141.34; 141.24; 140.79; 117.49; 107.49; 102.79; 100.24; 72.95; 70.92-68.73; 65.68; 41.45.

The monomer 7 was obtained similarly by replacing the 4’-aminobenzo-15-crown-5 with the 4’-aminobenzo-18-crown-6.

Synthesis of Monomer 8

2'-aminomethyl-3,4-ethylene-dioxythiophene (0.350 g; 2.04 mmol, leq.) And 2- (chloromethyl) pyridine hydrochloride (0.714g; 4.09 mmol, 2eq .) were added to a reactor containing 15 mL of water. The latter was heated to 60 ° C in an oil bath. 2.5 ml of a 5N sodium hydroxide solution was added to the reactor and the reaction mixture was allowed to stir for one hour. After one hour, 2- (chloromethyl) pyridine hydrochloride (0.335 g; 2.04 mmol, leq.) Dissolved in 5 ml of water was added and allowed to stir for 2 hours. After returning to room temperature, the mixture was extracted with diethyl ether (3x20 mL). After drying the combined organic phases with magnesium sulfate and evaporating the solvent, a pink oil was obtained. After purification by liquid chromatography on alumina as stationary phase and the mixture of solvents dichloromethane / methanol (99: 1) as mobile phase, the pure product 8 was obtained in the form of a yellow oil (0.126 g, Yield = 42%) .

Ή-NMR (400 MHz; CDC1 ³ ; 298K): ppm 8.44 (m, 2H); 7.55 (m, 2H); 7.38 (m, 2H); 7.06 (m, 2H); 6.19 (m, 2H); 4.20 (m, 1H); 4.10-3.90 (m, 2H); 3.85 (m, 4H); 2.81 (m, 2H). ¹³ C-NMR (75 or 150 MHz; CDC1 ₃ ; 298K): ô / ppm: 159.0; 149.1; 141.6; 136.4; 123.1; 122.2; 73.3; 72.3; 67.1; 67.0; 61.1; 55.1; 53.9; 49.2.

Synthesis of Monomers 9 and 10

A 100 ml three-necked flask, flamed and dried beforehand, was loaded with chloromethyl-EDOT (0.5 g; 2.62 mmol; 1.0 eq.), Hydroxy-methyl-18 crown 6 (0.526 g; 2.62 mmol; 1.0 eq.), potassium carbonate (0.725 g; 5.25 mmol; 2.0 eq.) and a catalytic amount of potassium iodide (0.043 g; 0.26 mmol; 0.1 eq.). The reaction medium was dissolved in DML (40 mL). This solution was stirred intensely at 100 ° C for 24 hours under inert conditions. Then, an equivalent of chloromethyl-EDOT was added and the reaction was continued for 24 hours under the same conditions. The reaction mixture was cooled to room temperature, then it was extracted with dichloromethane (3x20 mL). The combined organic fractions were washed with water (20 mL) and dried with magnesium sulfate. The solvent was evaporated in vacuo using a rotary evaporator and the product was purified by chromatography on an alumina column as a stationary phase and with a mixture of the solvents dichloromethane / ethyl acetate / methanol (1: 1: 1 ). A caramel-colored oil 10 (0.84 g; Yield = 72%) was thus obtained.

Ή-NMR (CDC1 ₃ , 400 MHz): ô (ppm): 6.33 (s, 2H); 4.23 (t, 2H); 4.11 (dd, 1H); 3.85 (m, 4H); 3.67 (m, 15H). ¹³ C-NMR (CDC1 ₃ , 150 MHz): ô (ppm): 141.64, 141.34; 140.89; 100.34; 92.83; 77.23; 74.22; 73.06; 70.8; 65.96; 65.78; 65.49; 61.74; 41.54.

The monomer 9 was synthesized in a similar way by replacing hydroxy-methyl-18-crown-6 with hydroxy-methyl-15-crown-5.

Example 2: Synthesis of polymers according to the invention

Example 2.1: Copolymerization of a monomer according to the invention and of STFSI

The monomers 1 to 3 were used for the synthesis of new electrolyte polymers by radical polymerization methods (via RAET) known in the literature. Different series of copolymers have been systematically synthesized, changing the ratio between the monomers according to the invention and the more conventional styrene (trifluoromethanesulfonyl) imide (STFSI) monomer providing solubility in water. The experimental procedure for the synthesis of the various electrolyte polymers is described below:

The necessary quantities of the STFSI monomer and of the monomer according to the invention were added to the schlenk type reactor as well as the adequate quantity of the chain transfer agent (CTA) and of the azobisisobutyronitrile radical initiator (AIBN) and solubilized with stirring in DMF to obtain a fairly concentrated reaction mixture. After several freeze-thaw cycles, the polymerization was carried out at 65 ° C. under an inert atmosphere for a period ranging from a few days to a few weeks depending on the desired molecular mass. The polymer was obtained after precipitation in tetrahydrofuran (THF) or THF / diethyl ether mixtures, then filtration, washing with THF and drying under vacuum at 65 ° C for at least one day.

The different ratios used for the monomers 1 (noted SP15Cr5SI) and 2 (noted SP18Cr6SI) are listed in table 1 below.

[0135] [Tables 1]

% by mass STFSI (ml) /% by mass SP15Cr5SI (m2) n _m i / n m2 Mw / K Da D % mass STFSI (ml) /% mass SP18Cr6SI (m2) n _m i / n m2 Mw / KD at D 60/40 1.4 154 2.8 50/50 1.4 96 2.3 70/30 3 128 2.2 60/40 2.1 44 1.4 80/20 5 115 2.2 70/30 3.3 144 2.3 85/15 7.2 130 2.4 85/15 7.9 135 2.6 95/5 24.1 108 1.8 90/10 12.5 127 2.3

In Table 1, D denotes the dispersity of the polymer chains and was determined by steric exclusion chromatography (DMF, polystyrene standards). Mw denotes the molar mass and was determined by steric exclusion chromatography.

The copolymers involving the monomer 3 and the STFSI monomer were synthesized with proportions by mass of monomer 3 of 5%, 10%, 30%, 40% and 50%.

The ion specificity has been demonstrated for a copolymer comprising 85% of STFSI monomer and 15% of monomer 2 by mass. This copolymer was characterized by UV-visible spectroscopy in the presence of increasing concentrations of different alkali metal salts (NaCl and KC1). It has thus been demonstrated that the polymer comprising the 18-crown-6 group has a greater sensitivity to the K ⁺ cation, preserving the specificity observed in the 18-crown-6 monomer.

Example 2.2 .- Electrochemical synthesis of polymers and electrochemical characterization of monomers and polymers

The electrochemical experiments were carried out using an AUTOLAB potentiostat controlled by a computer using the "General Purpose Electrochemical Software" (GPES) software. An electrochemical cell, with a configuration of 3 electrodes, equipped with a large Platinum surface electrode (mesh) and an Ag / AgCl electrode (in a 3M KC1 solution) as reference electrode (Bioanalytical Systems) was used for all experiments. Vitreous carbon was used as the working electrode for the characterization of the monomers. Working electrodes composed of gold films (120 nm thick deposited by thermal evaporation on gold substrates) or ITO were used for electropolymerizations and the characterization of polymer films formed in organic and aqueous media. The acetonitrile was degassed in the cell before use and kept under an inert atmosphere throughout the electropolymerization. Before any experiment, the glassy carbon electrode was polished with alumina (thickness) to guarantee reproducibility. The potentiometry experiments were carried out with a PalmSense potentiostat (Netherlands) controlled by the PS Trace 5.2 software in a cell with two electrodes.

The electrochemical profiles of the various monomers, as well as the conditions for polymerization and copolymerization with the EDOT monomer have been developed for obtaining ion-sensitive polymer films. The experimental procedure is described below.

An electrochemical cell with a configuration of 3 electrodes as mentioned before was used. A 0.1 M solution of lithium perchlorate in acetonitrile was used as the electrolyte in which the monomer (5 mM) to be characterized or polymerized was added. In the case of copolymerizations, the two monomers were added in the concentrations necessary to study the molar ratios, that is to have a total concentration of 10 mM. The characterization of the electrochemical profile of the commercial EDOT monomer and of the monomer 6 according to the invention was made using the glassy carbon electrode, previously polished to guarantee the use of a homogeneous and reproducible surface between the different experiments. To characterize the oxidation potential of each monomer, a unique potentiodynamic voltammetry cycle was carried out. The experiment was carried out using 0 V as initial and final potential as well as 1.5 V as maximum cycle potential. The scanning was done at a speed of 25 mV7 min. For electro (co-) polymerizations, a gold electrode was used (120 nm thick film deposited by metallic evaporation on glass substrates).

The scanning was done between 0 and 1.4 V for oxidation and between 1.4 V and -0.6 V for reduction at the same speed of 25 mV / min for 10 cycles. The polymer films obtained by electropolymerization were first of all characterized in the same type of electrolyte (0.1 M of lithium perchlorate in acetonitrile). A second series of characterizations was carried out in an aqueous medium with saline solutions of different concentrations (lOmM-lOOmM NaCl, KC1 or ZnCl ₂ ) for the analysis of ionic sensitivity.

The following polymers were synthesized: polymer obtained by electropolymerization of monomer 6, copolymer of monomer 6 with EDOT, in molar ratios (EDOT / 6) of 1/1, 2/1, 3/1 and 2 / 3.

FIG. 5a shows the electrochemical characterization of the monomer 6 relative to that of the EDOT monomer. FIG. 5b shows the electrochemical characterization of the various copolymers of monomer 6 with EDOT, in the order of the curves (starting from the top at the level of 0.0 on the abscissa) with EDOT / monomer 6 ratios of 2/1, 3 / 1, 2/3 and 1/1. FIG. 6a shows an image of scanning electron microscopy of the EDOT / monomer 6 copolymer with a 3/1 ratio. Figure 6b shows a scanning electron microscopy image of a 20 micron film formed by electropolymerization of monomer 6.

Example 3: Preparation of inks according to the invention

The electrolyte polymers obtained in Example 2.1 were used as stabilizers in aqueous dispersions of the EDOT monomer to produce, by oxidative polymerization, ion-sensitive inks based on PEDOT.

Three aqueous solutions of iron (III) chloride (0.036 mg; 5.84 mmol), ammonium persulfate (0.161 mg; 36.7 mmol) and of the copolymer obtained in Example 2.1 from the derivatives ion-sensitive styrene 1 to 3 with STESI (0.210 g - 0.420 g) were prepared and allowed to stir for 5 h before use. After this time, the solution of the electrolyte polymer was transferred to a reactor and the EDOT monomer was added (36 μL, 0.34 mmol) then allowed to stir for half an hour and under a flow of nitrogen at 10 ° C. Adding half the volumes of the oxidant solutions (iron chloride and ammonium persulfate) prepared beforehand enables the oxidative polymerization of EDOT to start. The reaction was left for 64 h under an inert atmosphere. The purification was carried out by ultrafiltration. A first wash was done with an IM solution of hydrochloric acid (150 mL). After concentrating the ink, it was further diluted with the same acid solution. After stirring for 2 hours, a third and final wash with MilliQ water was done. The ink was then filtered and analyzed by UV-Visible spectroscopy to determine a solution containing 1% dry matter according to our calibration. For their storage, the inks are protected from light and kept under magnetic stirring at constant room temperature.

FIG. 1 shows the synthesized inks and their comparison with the PEDOT reference system: PSS. FIG. 2 shows the electrochromic characterization of the dedoping time of an ink according to the invention and the characterization in electro-impedance spectroscopy of the same ink. The ink is a PEDOT ink: PSTESI 85: PSP15Cr5SI 15, SP15Cr5SI denoting monomer 1. The polymers and conductive complexes according to the invention can form stable aqueous dispersions. Their electrochromic properties are preserved. Their dedoping speed is comparable to that observed with the classic PEDOT: PSS complex.

The characterization of the polymers, conductive complexes and inks according to the invention can be carried out by conventional methods such as electrochemical impedance spectroscopy (EIS) or electrochromic spectroscopy (ECS). Some properties of the polymers, complexes and inks according to the invention even exceed those of the PEDOT: PSS complex, for example their capacitance, a property useful in OECT type devices.

Example 4: Study of the cytotoxicity of inks

Example 4.1: Preliminary study of cytotoxicity

The cytotoxicity of these inks was evaluated in a preliminary way (Ligure 3) via the observation of the morphology of the cells of a clonal line of beta cells in the presence of a thin layer of a film of 3 inks according to the invention. The composition of the 3 inks is detailed in Table 2 below:

[0154] [Tables2]

Formulation Agitation = 2 weeks Reagent F6 F7 F8 V / pL of PEDOT: Polyelectrolyte te 1000 pL of PEDOT: PSS_95PS18Cr6-l_5 1000 pL of PEDOT: PSTFI_85 -PS18Cr6-l_15 1000 pL of PEDOT: PSTFI_85 -PS18Cr6-l_15 DMSO 50 pL 50 pL 50 pL GOPS 20 pL 10 pL 20 pL Zonyl 4% 10 pL 10 pL 10 pL

The films are 150 nm films deposited by centrifugation (spin coating). After two days of cell culture, no significant cytotoxicity was observed and cell growth was similar to that under control conditions.

Example 4.2: Study of cell death in the presence and absence of ink according to the invention

Clonal β cells (INS832-13) were cultured for 24 hours on reference coverslips (a and b) or on coverslips coated with an ink of the polymer studied (c and d) (in this case PEDOT: PSTFSI_85 / monomer 2_15, based in the same stabilizer characterized in UV). The effect of thapsigargine (drug inducing apoptosis) was evaluated on the cells cultured on a coverslip coated with the ink according to the invention (d) and compared to the effect of the same drug on the cells cultured on the reference slide (b). Figure 4 presents the evaluation of the cytotoxicity of the indicated components. The effect of thapsigargine on βclonal cells (line INS-832/13) serves as a positive control thanks to its known cytotoxic activity. The coverslips were then subjected to fluorochrome labeling of living cells (in green) and dead cells (in red). Merged images are shown in Figure 4.

Measuring apoptosis in response to a stimulus makes it possible to assess the ability of cells to function normally and without damage. A low rate of apoptosis is always observed during the growth of cells in culture medium, and it is comparable in the two cases studied (strips with and without ink according to the invention), which means that the growth of cells is normal and cell mortality is negligible.

Thapsigargine is a non-competitive inhibitor of the calcium ATPase pump of the endoplasmic reticulum SERCA, which increases the intracellular calcium concentration by blocking the cell's ability to pump calcium into the endoplasmic reticulum, causing cell death.

After adding thapsigargine, comparable cell growth was observed under the two culture conditions (coverslips with / without ink according to the invention). In both cases, the cell morphology is clearly affected by the presence of thapsigargine. Note the decrease in the number of cells and the appearance of red coloration under apoptotic condition (b, d) while the cells are more numerous and alive in the presence of thin layers of ink without thapsigargine.

The ratio (dead cells) / (dead cells + living cells) was calculated in the presence and absence of ink according to the invention (Fig. 4 (e)) and it is comparable in the two cases, this which demonstrates that the inks according to the invention are not cytotoxic under these conditions.

Example 5: Manufacture of organo-electrochemical transistors

Finally, various organo-electrochemical transistors (OECT) were manufactured with the inks synthesized in Example 3. FIG. 7 presents a diagram of manufacturing and characterization of an OECT according to the invention, FIG. 8 presents the characterization results of said transistors. The manufacturing was done by conventional methods of metallic evaporation and the use of masks for the definition of the electrodes. The ink was deposited by centrifugation (spin coating).

The characterization was carried out in an aqueous medium.

The physical characterization of the OECTs, presenting figures of merit in the same orders of magnitude as those described in the state of the art, made it possible to demonstrate the potential use of the materials according to the invention as mixed conductive materials and as active materials in OECT type devices. Indeed, obtaining a transfer curve (I _D in FIG. 6) with an initial conductive state (“on”) with a current of 8.7 mA (max I _D , left Y axis) and which passes at zero current, 0 mA, non-conductive state ("off") when the grid voltage is applied (at Vg = 0.6 - 0.8V; X axis) indicates that the material conducts electrons and ions, being well a "mixed driver". In addition, the maximum amplification, known as the maximum of transconductance (g _m ), is of the order of the values found in the state of the art for devices manufactured with PEDOT: PSS.

Of course, various other modifications can be made to the invention in the context of the appended claims.

Claims (1)

Claims [Claim 1] Use of at least one polymer comprising at least one unit of formula (I) [Chem. 14] + · | + L 1 B in which A is a polymerizable monomer, L is a spacer arm, and B is a chemical group capable of complexing or chelating at least one ion chosen from the group consisting of K ⁺ , Na ⁺ , Ca ²⁺ and Zn ²⁺ , in a method of coating an electrode or manufacturing an organo-electrochemical transistor, in which either the polymer comprising at least one unit of formula (I) is an electronically conductive polymer, or the polymer comprising at least a unit of formula (I) is non-electronic conductor and is used in the form of a mixture with another electronic conductive polymer. [Claim 2] Use of at least one polymer comprising at least one unit of formula (I) according to claim 1, in which A is chosen from the group consisting of styrene, styrene sulfonate, and 3,4-ethylenedioxy thiophene. [Claim 3] Use of at least one polymer comprising at least one unit of formula (I) according to claim 1 or 2, wherein the polymer comprising at least one unit of formula (I) is used in the form of an ink. [Claim 4] Conductive ink comprising at least one polymer comprising at least one unit of formula (I) [Chem. 15] - [- A— L 1 B in which A is a polymerizable monomer, L is a spacer arm, and B is a chemical group capable of complexing or chelating at least one ion chosen from the group consisting of K ⁺ , Na ⁺ , Ca ²⁺ and Zn ²⁺ , in which either the polymer comprising at least one unit of formula (I) is an electronic conductive polymer, or the polymer comprising at least one unit of formula (I) is non-electronic conductor and is in the form of a mixture with another electronic conductive polymer. [Claim 5] Organoelectrochemical transistor comprising as semiconductor film a film comprising at least one polymer comprising at least one unit of formula (I) [Chem. 16] —A— L 1 B in which A is a polymerizable monomer, L is a spacer arm, and B is a chemical group capable of complexing or chelating at least one ion chosen from the group consisting of K ⁺ , Na ⁺ , Ca ²⁺ and Zn ²⁺ , in which either the polymer comprising at least one unit of formula (I) is an electronically conductive polymer, or the polymer comprising at least one unit of formula (I) is non-electronically conductive and is in the form of a mixture with another electronic conductive polymer. [Claim 6] Use of an organo-electrochemical transistor according to claim 5 for the detection of at least one flux of an ion chosen from the group consisting of K ⁺ , Na ⁺ , Ca ²⁺ and Zn ²⁺ at the level of a cell or a set of cells. [Claim 7] Use of an organo-electrochemical transistor according to claim 5 for the detection of the presence and / or the quantification of at least one ion chosen from the group consisting of K ⁺ , Na ⁺ , Ca ²⁺ and Zn ²⁺ in a liquid sample from a biological organism, in an environmental liquid sample, or in a liquid sample taken during the monitoring of a chemical process. [Claim 8] Polymer comprising at least one unit of formula (I) [Chem. 17] -AT- L 1 B in which A is a polymerizable monomer, L is a spacer arm, and B is a chemical group capable of complexing or chelating at least one ion chosen from the group consisting of K ⁺ , Na ⁺ , Ca ²⁺ and Zn ²⁺ . [Claim 9] The polymer of claim 8, wherein the polymer is a copolymer further comprising at least one unit selected from the group consisting of EDOT units, styrene (trifluoromethanesulfonyl) imide units, styrene units, styrene sulfonate units, and units of formula (I) with A, L and / or B different from those of the first unit of formula (I). [Claim 10] Polymer according to claim 8 or 9, in which the unit of formula (I) is chosen from the group consisting of: - 4-vinylbenzyl (sulfonyl) -4 '- (benzo-15-crown-5) -ylamine 1, - 4-vinylbenzyl (sulfonyl) -4 '- (benzo-18-crown-8) -ylamine 2, - the N- (4-vinylbenzyl) (pyridin-2-yl) -N - ((pyridin-2-yl) methyl) methylamine 3, - 4-vinylbenzyl-4 '- (methyl-15-crown-5) -methylether 4, - 4-vinylbenzyl-4 '- (methyl-18-crown-8) -methylether 5, -the (2,3-dihydrothieno [3,4-b] [1,4] dioxin-2-yl) -4 '- (benzo-15-crown-5) -m ethylamine 6, -the (2,3-dihydrothieno [3,4-b] [1,4] dioxin-2-yl) -4 '- (benzo-18-crown-6) -m ethylamine 7, -the (2,3-dihydrothieno [3,4-b] [1,4] dioxin-2-yl) -N, N-bis ((pyridin-2-yl) methy l) methanamine 8, - the (2,3-dihydrothieno [3,4-b] [1,4] dioxin-2-yl) -2- (methyl-15-crown-5) -m ethyl ether 9, and - the (2,3-dihydrothieno [3,4-b] [1,4] dioxin-2-yl) -2- (methyl-18-crown-6) -m ethyl ether 10. [Claim 11] Monomer of formula (II) [Chem. 18] A — L — B in which A is a polymerizable monomer, L is a spacer arm, and B is a chemical group capable of complexing or chelating at least one ion chosen from the group consisting of K ⁺ , Na ⁺ , Ca ^{2 +} and Zn ²⁺ , in which the monomer is chosen from the group consisting of: - 4-vinylbenzyl (sulfonyl) -4 '- (benzo-15-crown-5) -ylamine 1, - 4-vinylbenzyl (sulfonyl) -4' - ( benzo-18-crown-8) -ylamine 2, - 4-vinylbenzyl-4 '- (methyl-15-crown-5) -methylether 4, - 4-vinylbenzyl-4' - (methyl-18-crown- 8) -methylether 5, -la (2,3-dihydrothieno [3,4-b] [1,4] dioxin-2-yl) -4 '- (benzo-15-crown-5) -m ethylamine 6, -la (2,3-dihydrothieno [3,4-b] [1,4] dioxin-2-yl) -4 '- (benzo-18-crown-6) -m ethylamine 7, -la (2,3-dihydrothieno [3,4-b] [1,4] dioxin-2-yl) -N, N-bis ((pyridin-2-yl) methy l) methanamine 8, - (2,3-dihydrothieno [3,4-b] [1,4] dioxin-2-yl) -2- (methyl-15-crown-5) -m ethyl ether 9, and - (2,3-dihydrothieno [3,4-b] [1,4] dioxin-2-yl) -2- (methyl-18-crown-6) -m ethyl ether 10.